Asexual propagation for plants has been shown for some species to yield large numbers of genetically identical embryos, each having the capacity to develop into a normal plant. Such embryos must usually be further cultured under laboratory conditions until they reach an autotrophic “seedling” state characterized by an ability to produce their own food via photosynthesis, resist desiccation, produce roots able to penetrate soil and fend off soil microorganisms. Some researchers have experimented with the production of artificial seeds, known as manufactured seeds, in which individual plant somatic or zygotic embryos are encapsulated in a seed coat. Examples of such manufactured seeds are disclosed in U.S. Pat. No. 5,701,699, issued to Carlson et al., the disclosure of which is hereby expressly incorporated by reference.
Typical manufactured seeds include a seed coat, synthetic gametophyte and a plant embryo. The seed coat typically is a cylindrical capsule having a closed end and an open end. The synthetic gametophyte is placed within the seed coat, such that the gametophyte substantially fills the interior of the seed coat. A longitudinally extending hard porous insert, commonly known as a cotyledon restraint, may be centrally located within the synthetic gametophyte and includes a centrally located cavity extending partially through the length of the cotyledon restraint. The cavity is sized to receive the plant embryo therein. The well known plant embryo includes a radicle end and a cotyledon end. The plant embryo is deposited within the cavity of the cotyledon restraint cotyledon end first. The plant embryo is then sealed within the seed coat by at least one end seal. There is a weakened spot in the end seal to allow the radicle end of the embryo to penetrate the end seal.
Typically, the end seal is formed from a sheet of polymer material. In the past, a mechanical rod is used to stretch and, therefore, thin or weaken a small spot in the center of the seal. The weakened spot in the end seal is desirable because as the embryo begins to grow, the weakened spot permits tangential distribution of load to assist in the breakthrough of the embryo through the end seal.
A disk of predetermined diameter is then cut from the polymer sheet with the weakened spot in the center of the disk forming the end seal. After the end seals are cut from the polymer sheet, they are pushed out of a punch cavity with a rod and then fall by gravity on top of the seed coat. The seed coat with the end seal is then transferred to a separate station to heat fuse the end seals to the seed coats. Although such a process is effective at forming and sealing end seals to the seed coat of a manufactured seed, it is not without its problems.
First, it is difficult to maintain the exact location of the opening into the cavities of the cotyledon restraint as the manufactured seed passes through the production line. Second, the placement of the end seal over the seed coat is not reliable. Additionally, the end seals tend to stick to the punch or punch cavity. Further, fusing of the end seals to the seed coats is difficult and time consuming because the end seals must be heated to a point where the end seal is fused to the seed coat without damaging the dome or weakened area itself. Sometimes the fusing process melts holes in the weakened spot. The fusing process may cause damage to the plant embryo itself. Also, because of how the material was stretched, the resulting thinned or weakened area is not uniform in thickness. As a result, mass production of manufactured seeds is not only time consuming but also expensive.
Thus, there exists a need for a method of manufacturing and attaching an end seal to a seed coat of a manufactured seed that can produce and attach a large number of end seals to a manufactured seed at a relatively low cost, with a high degree of reliability, and without damaging the plant embryo located within the seed coat.